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Coated Vesicles: Vesicles formed when cell-membrane coated pits (Coated pits, Cell-membrane) invaginate and pinch off. The outer surface of these vesicles are covered with a lattice-like network of coat proteins, such as Clathrin, coat protein complex proteins, or Caveolins.

An Introduction to Endocytosis and Exocytosis

JoVE 5646

Cells can take in substances from the extracellular environment by endocytosis and actively release molecules into it by exocytosis. Such processes involve lipid membrane-bound sacs called vesicles. Knowledge of the molecular architecture and mechanisms of both is key to understanding normal cell physiology, as well as the disease states that arise when they become defective. This video will first briefly review a few pivotal discoveries in the history of endo- and exocytosis research. Next, some key questions will be examined, followed by a discussion of the prominent methods used to investigate these problems, including cell labeling, fusion assays, and fluorescence imaging. Finally, it will explore current research being conducted by scientists in the field today.


 Cell Biology

An Introduction to Transfection

JoVE 5068

Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery, whereas insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for transfected RNA is the cytoplasm, DNA must be transported to the nucleus for effective transfection. There, the DNA can be transiently expressed for a short period of time, or become incorporated into the genomic DNA, where the change is passed on from cell to cell as it divides. This video describes the basics behind chemical mediated transfections and introduces some of the most commonly-used reagents, including charged lipids, polymers, and calcium phosphate. Each step is described from the preparation of cells for transfection through analysis of transfection efficiency. Additionally, the applications section of this video-article describes the use of electroporation and a biolistic transfection as alternative methods for introducing nucleic acid into mammalian cells. It also describes an advanced use of transfection where co-transfection of interfering RNA and DNA are introduced as a way to down-regulate a naturally occurring protein while at the same time producing a


 Basic Methods in Cellular and Molecular Biology

Live Cell Imaging and 3D Analysis of Angiotensin Receptor Type 1a Trafficking in Transfected Human Embryonic Kidney Cells Using Confocal Microscopy

1Department of Biochemistry, Georgetown University Medical Center, 2Department of Medicine, Georgetown University Medical Center, 3Department of Physics, Georgetown University Medical Center, 4Department of Oncology, Georgetown University Medical Center

JoVE 55177


 Biology

An Unbiased Approach of Sampling TEM Sections in Neuroscience

1Department of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of Graz, 2Department of Pharmacology, Otto Loewi Research Center, Medical University of Graz, 3Division of Neurogeriatrics, Department of Neurology, Medical University of Graz, 4Institute of Pathology, Medical University of Graz, 5Department of Pathology, Medical Faculty, Otto von Guericke University Magdeburg

Video Coming Soon

JoVE 58745


 JoVE In-Press

Pulling Membrane Nanotubes from Giant Unilamellar Vesicles

1Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, CNRS UMR168, 2Department of Genetics and Complex Diseases, T. H. Chan School of Public Health, Harvard Medical School, 3Department of Cell Biology, Harvard Medical School, 4Sorbonne Universités, UPMC University Paris 06, 5Center for Studies in Physics and Biology, The Rockefeller University

JoVE 56086


 Biology

Spontaneous Formation and Rearrangement of Artificial Lipid Nanotube Networks As a Bottom-Up Model for Endoplasmic Reticulum

1Centre for Molecular Medicine Norway, Faculty of Medicine, University of Oslo, 2Department of Chemistry, Faculty of Mathematics and Natural Sciences, University of Oslo, 3Department of Chemistry and Chemical Engineering, Chalmers University of Technology

Video Coming Soon

JoVE 58923


 JoVE In-Press

Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies

1Institut Curie, Centre de Recherche, CNRS, UMR 168, PhysicoChimie Curie, Université Pierre et Marie Curie, 2Kavli Institute for Brain and Mind, University of California, San Diego, 3Molecular Physiology and Biophysics Section, National Institute for Neurological Disorders and Stroke, National Institute of Health

JoVE 52281


 Biology

A Unified Methodological Framework for Vestibular Schwannoma Research

1Eaton Peabody Laboratories, Department of Otolaryngology, Massachusetts Eye and Ear, 2Department of Otolaryngology, Harvard Medical School, 3Department of Otolaryngology, Vienna General Hospital, Medical University of Vienna, 4Program in Speech and Hearing Bioscience and Technology, Harvard Medical School

JoVE 55827


 Cancer Research

Efficient Gene Delivery into Multiple CNS Territories Using In Utero Electroporation

1Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children’s Hospital Research Institute, University of Calgary, 2Department of Medical Genetics, Alberta Children’s Hospital Research Institute, Hotchkiss Brain Institute, University of Calgary

JoVE 2957


 Neuroscience

Biodistribution of Nano-drug Carriers: Applications of SEM

JoVE 10472

Source: Peiman Shahbeigi and Sina Shahbazmohamadi, Biomedical Engineering Department, University of Connecticut, Storrs, Connecticut

Nanoparticles have been increasingly used research towards targeted drug delivery and controlled drug release. While most of these particles have been developed as polymeric or liposomal particles because of their biocompatibility, there is a trend in current research toward the use of metallic and magnetic nanoparticles. These metallic nanoparticles were originally used as a contrast agent in imaging, but recent advances have shown how important they could be in drug and gene delivery and in therapeutics. Gold, silver, and paramagnetic nanoparticles have the greatest share in research being done. They have been shown to have good biocompatibility and certain varieties of magnetic nanoparticles have already been developed and distributed as therapeutic targeted drugs.   These heavy elements are typically imaged for research using fluorescence to evaluate delivery and distribution, but their atomic weights are good qualifications for increased contrast in backscatter electron analysis using a scanning electron microscope (SEM). Energy dispersive X-ray spectroscopy, which uses characteristic X-rays emitted upon electron beam interac


 Biomedical Engineering

An Enzyme- and Serum-free Neural Stem Cell Culture Model for EMT Investigation Suited for Drug Discovery

1Dept. of Biomedicine, Pharmacenter, University of Basel, 2Molecular Signalling and Gene Therapy, Narayana Nethralaya Foundation, Narayana Health City, 3Brain Ischemia and Regeneration, Department of Biomedicine, University Hospital Basel, 4Department of Neurosurgery, Klinikum Idar-Oberstein, 5Department of Neurosurgery and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, 6Department of Neurology, Laboratory of Molecular Neuro Oncology, University Hospital of Zurich

JoVE 54018


 Developmental Biology

SNARE-mediated Fusion of Single Proteoliposomes with Tethered Supported Bilayers in a Microfluidic Flow Cell Monitored by Polarized TIRF Microscopy

1Department of Cellular and Molecular Physiology, Yale University School of Medicine, 2Nanobiology Institute, Yale University, 3Department of Molecular Biophysics and Biochemistry, Yale University, 4Laboratoire de Neurophotonique, Université Paris Descartes, Faculté des Sciences Fondamentales et Biomédicales, Centre National de la Recherche Scientifique (CNRS)

JoVE 54349


 Neuroscience

A Cell-based Assay to Investigate Non-muscle Myosin II Contractility via the Folded-gastrulation Signaling Pathway in Drosophila S2R+ Cells

1Department of Biology, The University of North Carolina at Chapel Hill, 2Department of Biology, Reed College, 3Carolina Center for Genome Sciences, The University of North Carolina at Chapel Hill, 4Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill

JoVE 58325


 Developmental Biology

Polyethyleneimine-coated Iron Oxide Nanoparticles As a Vehicle for the Delivery of Small Interfering RNA to Macrophages In Vitro and In Vivo

1Key Laboratory of Ministry of Education for Developmental Genes & Human Diseases, Institute of Life Sciences, Southeast University, 2State Key Laboratory of Bioelectronics, Jiangsu Key Laboratory for Biomaterials & Devices, School of Biological Science & Medical Engineering, Southeast University, 3Key Laboratory of Ministry of Education for Developmental Genes & Human Diseases, Medical School, Southeast University

Video Coming Soon

JoVE 58660


 JoVE In-Press

LDL Cholesterol Uptake Assay Using Live Cell Imaging Analysis with Cell Health Monitoring

1Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, 2Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, 3Department of Molecular and Cellular Pharmacology, University of Miami Leonard M. Miller School of Medicine, 4Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Leonard M. Miller School of Medicine, 5John P. Hussman Institute for Human Genomics, University of Miami Leonard M. Miller School of Medicine, 6Department of Medicine, Division of Endocrinology, Metabolism and Endocrinology and the Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, 7Vascular Biology Institute, University of Miami Leonard M. Miller School of Medicine, 8Peggy and Harold Katz Family Drug Discovery Center, University of Miami Leonard M. Miller School of Medicine

Video Coming Soon

JoVE 58564


 JoVE In-Press

Generation of Cationic Nanoliposomes for the Efficient Delivery of In Vitro Transcribed Messenger RNA

1Department of Thoracic and Cardiovascular Surgery, Clinical Research Laboratory, University Medical Center, 2Atherothrombosis and Vascular Biology, Baker Heart & Diabetes Institute, 3Department of Medicine, Monash University

Video Coming Soon

JoVE 58444


 JoVE In-Press

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator

1Institute of Medical Biochemistry, Center for Molecular Biology of Inflammation, University of Münster, 2Institute of Biochemistry, University of Münster, 3Cluster of Excellence 'Cells in Motion', University of Münster

Video Coming Soon

JoVE 58224


 JoVE In-Press

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